Molecular codes and in vitro generation of hypocretin and melanin concentrating hormone neurons.
Ali SeifinejadSha LiCyril MikhailAnne VassalliSylvain PradervandYoan ArribatHassan Pezeshgi ModarresBridget AllenRosalind M JohnFrancesca AmatiMehdi TaftiPublished in: Proceedings of the National Academy of Sciences of the United States of America (2019)
Hypocretin/orexin (HCRT) and melanin concentrating hormone (MCH) neuropeptides are exclusively produced by the lateral hypothalamus and play important roles in sleep, metabolism, reward, and motivation. Loss of HCRT (ligands or receptors) causes the sleep disorder narcolepsy with cataplexy in humans and in animal models. How these neuropeptides are produced and involved in diverse functions remain unknown. Here, we developed methods to sort and purify HCRT and MCH neurons from the mouse late embryonic hypothalamus. RNA sequencing revealed key factors of fate determination for HCRT (Peg3, Ahr1, Six6, Nr2f2, and Prrx1) and MCH (Lmx1, Gbx2, and Peg3) neurons. Loss of Peg3 in mice significantly reduces HCRT and MCH cell numbers, while knock-down of a Peg3 ortholog in zebrafish completely abolishes their expression, resulting in a 2-fold increase in sleep amount. We also found that loss of HCRT neurons in Hcrt-ataxin-3 mice results in a specific 50% decrease in another orexigenic neuropeptide, QRFP, that might explain the metabolic syndrome in narcolepsy. The transcriptome results were used to develop protocols for the production of HCRT and MCH neurons from induced pluripotent stem cells and ascorbic acid was found necessary for HCRT and BMP7 for MCH cell differentiation. Our results provide a platform to understand the development and expression of HCRT and MCH and their multiple functions in health and disease.
Keyphrases
- spinal cord
- single cell
- drug delivery
- metabolic syndrome
- poor prognosis
- physical activity
- healthcare
- public health
- induced pluripotent stem cells
- type diabetes
- sleep quality
- mental health
- adipose tissue
- risk assessment
- gene expression
- stem cells
- binding protein
- genome wide
- dna methylation
- bone marrow
- high resolution
- health information
- liquid chromatography
- uric acid
- skeletal muscle
- human health
- health promotion